Aptamer-Based Detection of Cardiac Biomarker Glycosylation States Using APT-SNAP

DESCRIPTION provided by applicant Aptamer Based Detection of Cardiac Biomarker Glycosylation States Using APT SNAP Arrays PIs Christopher L Warren and Mary S Ozers The presence of cardiac related biomarkers in patient biospecimens can provide important insight into diagnosis of cardiovascular disease its progression and optimal therapies for treatment Most of the cardiac biomarkers in clinical use detect heart failure and the cardiovascular field is in great need of biomarkers that span the cardiovascular disease spectrum from developing atherosclerosis to late stage disease The clinically accepted biomarker for heart failure BNP is modified by glycosylation and the glycosylated form of BNP predominates in patients with heart failure and chronic renal failure However the leading diagnostic assay for BNP does not detect the glycosylated forms Furthermore robust technologies to accurately detect multiple biomarkers especially glycosylated or other modified forms of biomarkers are lacking To address this significant diagnostic need for heart disease patients we are developing the APT SNAP Aptamer Specificity and Affinity for Proteins microarray which displays millions of DNA and RNA aptamers as a prototype high throughput device to design discover and optimize high affinity aptamers for heart disease biomarkers Nucleic acid aptamers are composed of either DNA or RNA and can adopt a compact three dimensional structure that recognizes a target with exceptional specificity and high affinity making them useful as a sensor of cardiac biomarkers In this proposal aptamers will be identified on the APT SNAP array for six major cardiac biomarkers as proof of principle A key invention of the proposal is a novel methodology to synthesize high density RNA aptamers arrays Identified aptamers for the cardiac biomarkers will be tested for their limit of detection n human serum The APT SNAP array will identify aptamers that recognize distinct glycosylation structures at specific residues of a biomarker using NT proBNP as a model biomarker To accomplish this we will synthesize a set of NT proBNP proteins each with a distinct glycosylation pattern and iteratively design aptamers that specifically recognize the glycosylated BNP forms To expand the universality of the assay beyond the limitations of antibodies currently used in many clinical assays a fluorescent protein dye will be used to correlate array fluorescence intensity to biomarker concentration The Specific Aims of this glycomics proteomics based proposal are Design novel high density DNA and RNA aptamer microarrays to identify aptamers for cardiac biomarkers Develop a panel of aptamers to detect specific glycosylated forms of cardiac biomarkers The ultimate goals are to develop APT SNAP as a biomarker discovery platform Phase I clinically correlate additional biomarkers Phase II and develop an affordable annual point of care diagnostic test Phase IIB for hundreds of cardiac biomarkers for physician use These technologies will impact the ability to diagnose heart disease accurately allowing for better treatment options improved patient care and longer lifespans PUBLIC HEALTH RELEVANCE Aptamer Based Detection of Cardiac Biomarker Glycosylation States Using APT SNAP Arrays PIs Christopher L Warren and Mary S Ozers The ability to check patients annually for heart disease with a cost effective assay that simultaneously detects hundreds of cardiac biomarkers including specific glycosylated forms would transform cardiovascular medicine Nucleic acid aptamers are composed of either DNA or RNA and can adopt a compact three dimensional structure that recognizes a target with exceptional specificity and affinity making them useful as a sensor of cardiac biomarkers This proposal will develop the APT SNAP Aptamer Specificity and Affinity for Protein array technology to design and optimize highly specific aptamers that recognize glycosylated forms of cardiac biomarkers with the goal of creating an affordable diagnostic test of heart disease for point of care use by physicians